Fig 1a: LB & Ampicillin plate with DNA. Colonies of BL21(DE3) bacteria transformed with the plasmid pGEM-gbr22 that grew after being incubated at 37 degrees Celsius for 15 hours.
Fig 1b: LB & Ampicillin plate with No DNA. There are no colonies on the control plate after being incubated at 37 degrees Celsius for 15 hours.
Fig 1c: No ampicillin in the agar. Ample bacterial growth (numerous colonies) can be seen on the Fun Plate. The plate was divided in half for Sanjna Z. and Sri N. to collect samples from Painter 2.14. The floor, counters, and cabinet tops were swabbed generously. The plate was incubated for about 15 hours at 37 degrees Celsius.
Fig 2: A single colony was scooped from the LB & Ampicillin plate with DNA that had been transformed with the plasmid pGEM-gbr22. The large purple culture in the flask grew after about 15.5 hours in the shaking incubator at 37 degrees Celsius at a rate of 250 rpm.
Fig 3: Large culture of BL21(DE3) bacteria transformed with the plasmid DNA pGEM-gbr22 centrifuged down to a wet pellet for 10 minutes at 5,000 rmp at 4 degrees Celsius. Wet pellet weight #1= 0.32 g. Wet pellet weight #2= 0.20 g.
Fig 4: 5 mL of purple solution in the Elution 1 tube. After pouring 5 mL of elution buffer (1x PBS and 250 mM imidazole), the pGEM-gbr22 protein separated from the Ni-NTA resin and flowed into the tube.
Fig 5: 55 mL of a relatively clear solution in the Elution 2 tube. A sample of this liquid was taken (Sample 6) to be tested in the Protein Characterization (Part III) of this lab.
Fig 6: Wet protein gel. Ladder in first well. 1 = James F. Sample 2. 2 = James F. Sample 3. 3 = blank. 4 = James F. Sample 4. 5 = blank. 6 = James F. Sample 5. 7 = James F. Sample 6. 9 = Sanjna Z. Sample 5. Gel electrophoresis machine would not stay on long enough to execute the process to completion.
Fig 7: Dry gel after 1.5 hours of drying at 75 degrees Celsius. Sample covered with cellophane and placed on Whatman paper. Location and thickness of lines for each Sample 5 indicative of protein purity. Additional methods of purification still needed to achieve an ideal sample.
Introduction:
Materials & Methods:
Results:
Discussion:
Conclusions:
References: